scholarly journals An investigation of the potential effect of sperm nuclear vacuoles in human spermatozoa on DNA fragmentation using a neutral and alkaline Comet assay

Andrology ◽  
2017 ◽  
Vol 5 (2) ◽  
pp. 392-398 ◽  
Author(s):  
E. Pastuszek ◽  
J. Kiewisz ◽  
P. Skowronska ◽  
J. Liss ◽  
M. Lukaszuk ◽  
...  
Keyword(s):  
Comet Assay ◽  
Dna Fragmentation ◽  
Potential Effect ◽  
Human Spermatozoa ◽  
Alkaline Comet Assay ◽  
Sperm Nuclear Vacuoles

Dimeric NGF Mimetic Attenuates Hyperglycaemia and DNA Damage in Mice with Streptozotocin-Induced Early-Stage Diabetes

2020 ◽  
Vol 20 (3) ◽  
pp. 453-463
Author(s):  
Svetlana Yagubova ◽  
Aliy Zhanataev ◽  
Rita Ostrovskaya ◽  
Еlena Anisina ◽  
Тatiana Gudasheva ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Early Stage ◽  
Alkaline Comet Assay ◽  
Β Cells ◽  
Ghost Cells ◽  
Low Weight

Background: NGF deficiency is one of the reasons for reduced β-cells survival in diabetes. Our previous experiments revealed the ability of low-weight NGF mimetic, GK-2, to reduce hyperglycaemia in a model of advanced diabetes. The increase in DNA damage in advanced diabetes was repeatedly reported, while there were no data about DNA damage in the initial diabetes. Aim: The study aimed to establish whether DNA damage occurs in initial diabetes and whether GK-2 is able to overcome the damage. Methods: The early-stage diabetes was modelled in Balb/c mice by streptozotocin (STZ) (130 mg/kg, i.p.). GK-2 was administered at a dose of 0.5 mg/kg, i.p., subchronically. The evaluation of DNA damage was performed using the alkaline comet assay; the percentage of DNA in the tail (%TDNA) and the percentage of the atypical DNA comets (“ghost cells”) were determined. Results: STZ at this subthreshold dose produced a slight increase in glycemia and MDA. Meanwhile, pronounced DNA damage was observed, concerning mostly the percentage of “ghost cells” in the pancreas, the liver and kidneys. GK-2 attenuated the degree of hyperglycaemia and reduced the % of “ghost cells” and %TDNA in all the organs examined; this effect continued after discontinuation of the therapy. Conclusion: Early-stage diabetes is accompanied by DNA damage, manifested by the increase of “ghost cells” percentage. The severity of these changes significantly exceeds the degree of hyperglycaemia and MDA accumulation. GK-2 exerts an antihyperglycaemic effect and attenuates the degree of DNA damage. Our results indicate that the comet assay is a highly informative method for search of antidiabetic medicines.

scholarly journals Potential effect of tobacco cigarettes smoking on global DNA methylation status and protamines transcripts in human spermatozoa

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Mohammed M. Laqqan ◽  
Maged M. Yassin
Keyword(s):  
Dna Methylation ◽  
Cigarette Smoking ◽  
Dna Fragmentation ◽  
Methylation Status ◽  
Human Spermatozoa ◽  
Global Dna Methylation ◽  
Transcription Level ◽  
Heavy Smokers ◽  
Protamine 1 ◽  
Protamine 2

Abstract Background Epigenetics refers to an alteration in gene expression without alteration in the sequence of DNA and this process may be affected by environmental factors and lifestyle like cigarette smoking. This study was designed to evaluate the potential effect of cigarette smoking on the global DNA methylation status and the transcription level of protamine 1 and protamine 2 in human spermatozoa. A total of 188 semen samples were collected from men with a mean age of 34.9 ± 5.8 years old (98 heavy smokers and 90 non-smokers). The DNA and RNA were isolated from purified spermatozoa, then the status of global DNA methylation and the transcription level of protamine 1 and protamine 2 were evaluated using ELISA and qPCR, respectively. The chromatin non-condensation and DNA fragmentation in human spermatozoa were evaluated using chromomycin A3 staining and TUNEL assay, respectively. Results A significant increase has been found in the status of global DNA methylation in spermatozoa of heavy smokers compared to non-smokers (7.69 ± 0.69 ng/μl vs. 4.90 ± 0.40 ng/μl, P < 0.001). Additionally, a significant reduction has been found in transcription level of protamine 1 (25.49 ± 0.31 vs. 23.94 ± 0.40, P < 0.001) and protamine 2 (28.27 ± 0.39 vs. 23.45 ± 0.30, P < 0.001) in heavy smokers. A downregulation has been found in the transcription level of protamine 1 and protamine 2 with a fold change of 0.497 and 0.047, respectively. A significant increase has been shown in the level of DNA fragmentation and chromatin non-condensation in heavy smokers compared to non-smokers (P < 0.001). On the other hand, a significant positive correlation has been found between sperm chromatin non-condensation, sperm DNA fragmentation, transcription level of protamine 1, transcription level of protamine 2, and global DNA methylation status (r = 0.304, P < 0.001; r = 0.399, P < 0.001; r = 0.216, P = 0.003; r = 0.494, P < 0.001, respectively). Conclusion Tobacco cigarette smoking has a potential influence on the global DNA methylation and the transcription level of protamine genes in human spermatozoa, and consequently, affect negatively on the semen parameters.

scholarly journals Analysis of DNA damage in cells excreted in urine of cervical cancer patients using alkaline comet assay

2014 ◽  
Vol 7 (Suppl 1) ◽  
pp. P12
Author(s):  
Avani Patel ◽  
Mihir Shah ◽  
Pinaki Patel ◽  
Trupti Patel
Keyword(s):  
Cervical Cancer ◽  
Dna Damage ◽  
Comet Assay ◽  
Cancer Patients ◽  
Alkaline Comet Assay ◽  
In Cells

Monitoring of DNA damage in environmentally boron exposed people in Turkey by the alkaline comet assay

2007 ◽  
Vol 172 ◽  
pp. S72
Author(s):  
Ulku Undeger ◽  
Mehmet Korkmaz ◽  
Sevtap Aydin ◽  
Nursen Basaran
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Alkaline Comet Assay

scholarly journals Measurements using the alkaline comet assay predict bladder cancer cell radiosensitivity

2003 ◽  
Vol 89 (12) ◽  
pp. 2271-2276 ◽  
Author(s):  
M A L Moneef ◽  
B T Sherwood ◽  
K J Bowman ◽  
R C Kockelbergh ◽  
R P Symonds ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Comet Assay ◽  
Cancer Cell ◽  
Bladder Cancer Cell ◽  
Alkaline Comet Assay
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